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Abstract Algae are found everywhere around us. In fact just about everywhere where there is a light to carry out photosynthesis. Algae grow in almost any aquatic environment and use light and carbon dioxide (CO2) to create biomass. Algae are not only a source of protein but also of other valuable materials such as fats, carbohydrates and pigments. This investigation was conducted aiming at the mass production of purified pigments of Chlorella vulgaris, Spirulina platensis and Nannochloropsis oculata. Chlorella vulgaris, Spirulina platensis and Nannochloropsis oculata were cultivated at indoor and outdoor conditions. For indoor cultivation: Incubation the used algae under BGII medium conditions. After cultivation period (19 days) of Chlorella vulgaris, the results showed that the dry weight was (0.92 g.l-1), degree of multiplication (1.93) and doubling time (0.99). Total chlorophyll content was (18.2 mg.l-1) with degree of multiplication (1.85) and doubling time (10.35). While, total carotene was (12.69 mg.l-1) with degree of multiplication (2.38) and doubling time (7.7). After cultivation period (12 days) of Spirulina platensis, the results showed that the dry weight was (1.44 g.l-1), degree of multiplication (2.26) and doubling time (3.47). Total chlorophyll content was (1.12 mg.l-1) with degree of multiplication (1.03) and doubling time (11.75). While, total carotene was (2.4 mg.l-1) with degree of multiplication (0.88) and doubling time (13.59). After cultivation period (13 days) of Nannochloropsis oculata, the results showed that the dry weight was(0.5g.l-1),degree of multiplication (4.64) and doubling time (2.77). Total chlorophyll content was (2.38 mgL-1) with degree of multiplication (2.28) and doubling time (5.78). While, total carotene was (0.85 mgL-1) with degree of multiplication (1.09) and doubling time (11.55). from the previous results Spirulina platensis surpasses all other used algae in dry weight. While in the outdoor cultivation, three units were used as follows: 1- Open plate photobioreactor: 1000L open plate was used for growth of Chlorella vulgaris. Cultivation was performed for 14 days and after that dry weight was in the 14th day (3.26 g.l-1) with degree of multiplication (3.35) and doubling time (4.33). Total chlorophyll was (31.35 mg.l-1) at the same day with degree of multiplication (2.3) and doubling time (6.86). But, total carotene was (32.87 mg.l-1) in the 14th day also with degree of multiplication (1.26) and doubling time (11.55). 2- Open pond: 15000L open pond was used for growth of Spirulina platensis. After 10 days of cultivation, dry weight was (1.09 g.l-1) with degree of multiplication (5.62) and doubling time (4.95). Total chlorophyll was (4.79 mg.l-1) at the same day with degree of multiplication (0.41) and doubling time (9.67). But, total carotene was (1.4 mg.l-1) in the end of cultivation period with degree of multiplication (0.61) and doubling time (6.66).3- Zigzag shape photobioreactor: 1000L zigzag shape was used for growth of Nannochloropsis oculata. After 9 days of cultivation, dry weight was (1.72 g.l-1) with degree of multiplication (1.38) and doubling time (6.3). Total chlorophyll was (2.37 mg.l-1) at the same day with degree of multiplication (1.84) and doubling time (4.95). But, total carotene was (4.3 mg.l-1) in the end of cultivation period with degree of multiplication (4.35) and doubling time (2.04). Spectrophotometrically, four solvents were used, dimethylsulfoxide, methanol, acetone and ethanol with wet and dry samples of different strains as follows: 1- Chlorella vulgaris: A- Chlorophyll a: It was found that dimethylsulfoxide has the highest pigment extraction efficiency of wet sample (27.73 mg/g). While, methanol was the best solvent for extraction of dry sample (44.29 mg/g). B- Chlorophyll b: It was found that dimethylsulfoxide was the better solvent for the chlorophyll extraction efficiency of both wet and dry samples (70.03, 38.12 mg/g) respectively. C- Total chlorophyll: Like chlorophyll b, dimethylsulfoxide was the dominant and best solvent with both wet and dry samples (97.76, 54.96 mg/g) respectively. D- Total carotene: Like wet samples of all above pigments, dimethylsulfoxide surpasses all other used solvents (24.3 mg/g). On contrast, ethanol with dry sample reached to maximum chlorophyll extraction efficiency (34.0 mg/g). 2- Spirulina platensis: For wet samples of all pigments of Spirulina platensis, the same results were obtained of chlorophyll a, c, total chlorophyll and total carotene (27.5, 63.8, 91.26 and 2.97) respectively with dimethylsulfoxide. For dry samples, the extraction efficiency of chlorophyll a, c and total chlorophyll were (39.67, 66.44 and 106.1 mg/g) respectively, with methanol. While, total carotene reached to maximum content (9.54 mg/g) with dimethylsulfoxide. The results that obtained for extraction of phycocyanin from both wet and dry sample were (2.03, 4.23 mg/g), respectively. 3- Nannochloropsis oculata: A- Chlorophyll a: It was found that acetone has the highest chlorophyll a extraction efficiency with both wet and dry samples (61.7 and 138.4 mg/g) respectively. B- Total carotene: For wet sample, dimethylsulfoxide was the best solvent (14.49 mg/g). Like dry sample of Chlorella vulgaris, ethanol has the highest carotene extraction efficiency (19.74 mg/g).Using HPLC method for determination of algal pigments from the three algal strains it was found that: For Chlorella vulgaris, acetone and methanol were excellent extraction solvents with wet and dry samples respectively. For wet samples, acetone was found (26.67, 34.8 mg/g) for both chlorophyll a and β-carotene respectively. While, for dry samples, methanol was found (44.97, 52.24 mg/g) for both chlorophyll a and β-carotene respectively. For Spirulina platensis, dimethylsulfoxide surpasses all other used solvents of all pigments and samples. For wet samples, it was found (25.13, 18.6 mg/g) for both chlorophyll a and β-carotene respectively. For dry samples, it was found (80.7, 75.8 mg/g) for both chlorophyll a and β- carotene respectively. For Nannochloropsis oculata, methanol surpasses all other used solvents of all pigments for wet samples. It was found (71.36, 32.25 mg/g) for both chlorophyll a and β-carotene respectively. For dry samples, methanol and ethanol were found (102.48, 48.51 mg/g) for both chlorophyll a and β- carotene respectively. The GC/MS analysis of the produced Cu-chlorophyll revealed some compounds. The Cu-chlorophyll extract was characterized by the presence of Quercetin-7,3’,4’- trimethoxy (39.64%), Lucenin 2 (31.93%), Isobutyl phthalate (56.35%), Palmitic acid, ethyl ester (61.21%), 1,1,2,2-Tetrachloroethane (37.76%), 2,5- Octadecadiynoic acid, methyl ester (8.74%) and Ether, bis (dichloromethyl) (4.38%), Hexadecadienoic acid, methyl ester (CAS) (5.38%), Linoleic acid ethyl ester(8.05%), 9 Hexadecenoic acid, ethyl ester (3.74%), Stearic acid, ethyl ester (3.21%), Octadecanoic acid, ethyl ester (CAS) (8.30%), Dibutyl phthalate (6.91%), Phthalic acid, bis(2-ethylhexyl) ester (6.23%), and Heptadecane (4.30%) as a major components. along with some other minor components presented in trace amounts. Na-Cu-Chlorophyllin was produced and the result of two peaks of Na-Cu-Chlorophyllin was appearing 3.88mg/g with retention time 8.416 min and the second peak was appearing 4.82mg/g with retention time 9.878 min. β-carotene crystallization was produced and the result of the chromatogram was characterized by peak at 10.933 min. The identity of the peak was confirmed by determination of relative retention time and by spiking with standard β- carotene. The result of β-carotene was 0.484 mg/g. Phycocyanin pigment was produced from Spirulina and it was found that the concentration of phycocyanin gave 4.8 mg/g and purity ratio gave 1.37%. |